JP7017749B2 - Anti-allergic agents and food compositions for the prevention or treatment of allergic diseases - Google Patents

Description

The present invention relates to an antiallergic agent. According to the antiallergic agent of the present invention, allergic diseases can be prevented, alleviated, or treated. The present invention also relates to a food composition for the prevention or treatment of allergic diseases.

Allergy means that the immune system responds to normally harmless antigens, resulting in adverse effects on the human body. Allergic diseases include atopic dermatitis, bronchial asthma, allergic rhinitis, pollinosis, and food allergies, and it is said that more than 30% of Japanese have some allergies.

Type I allergy represented by pollinosis is caused by IgE produced by B cells, which are a type of lymphocyte, and develops. In the case of hay fever, when pollen is recognized as an allergen (antigen) and IgE that specifically binds to it is produced in the body, IgE binds to the surface of basophils and mast cells via the IgE receptor (FcεRI). When pollen, which is an allergen, invades the body, it binds to IgE, which stimulates the transmission of signals into basophils and mast cells, and holds substances that cause allergic symptoms such as histamine. Granules are released from the cells. This phenomenon is called degranulation. Histamine released extracellularly by degranulation irritates the mucous membrane and causes allergic symptoms. Therefore, in order to prevent or treat allergic symptoms, it is important to suppress the release of granules by basophils and mast cells.

Various research activities are being carried out to control and treat allergies. Among them, ingredients derived from foods are excellent in safety, and various ingredients having an antiallergic effect have been reported. For example, Patent Document 1 describes an antiallergic agent containing apolactoferrin. Further, Patent Document 2 describes an antiallergic agent containing lees. However, the current situation is that a safe substance that has a sufficient desired antiallergic effect and can be actually used as a food or a pharmaceutical product has not yet been developed.

Japanese Unexamined Patent Publication No. 2013-236623 Japanese Unexamined Patent Publication No. 2009-292785

An object of the present invention is to provide a safe anti-allergic agent that can be ingested on a daily basis, has no fear of side effects, and can be expected to have excellent effects.

As a result of diligent studies on safe anti-allergic agents, the present inventors have surprisingly found that coriander extract exhibits an anti-allergic effect.
The present invention is based on these findings.
Therefore, the present invention
[1] An antiallergic agent containing a pulverized or extract of coriander,
[2] The antiallergic agent according to [1], which is an antiallergic agent for type I allergy.
[3] The antiallergic agent according to [1] or [2], wherein the crushed or extract of coriander is a crushed or extract of leaves.
[4] The antiallergic agent according to any one of [1] to [3], and [5] [1] to [4], wherein the extract is an aqueous solvent or an extract of a mixture of an aqueous solvent and an organic solvent. A food composition for preventing or treating allergic diseases, which comprises the antiallergic agent according to any one of [4] and foods and drinks.
Regarding.

According to the antiallergic agent of the present invention, the onset of allergic diseases can be prevented, or the symptoms thereof can be alleviated or treated. Furthermore, according to the antiallergic agent of the present invention, it is possible to provide foods and drinks having an antiallergic effect and having high safety by suppressing degranulation.

It is a figure which showed the method of preparing the extract from the fresh leaf of coriander (pakuchi) using the sodium phosphate buffer solution. It is a graph (A) which showed the inhibitory effect of the cilantro water-soluble extract on the degranulation of RBL-2H3 cells, and the graph (B) which examined the cytotoxicity of the cilantro leaf water-soluble extract. It is a graph which measured the change of the intracellular calcium ion concentration by the coriander leaf water-soluble extract (CLE). It is a graph which showed the inhibitory effect (A) of CLE on the degranulation by A23187 stimulation and the change (B) of the calcium ion concentration in detail. It is a graph which showed the inhibitory effect (A) of CLE on degranulation by Thapsigargin stimulation and the change (B) of the calcium ion concentration in detail. It is a photograph and a graph which examined the influence of the coriander leaf water-soluble extract on the degranulation signal transduction by Western blotting. It is a schematic diagram which showed the signal transduction of a degranulation in a cell. It is a figure which showed the experimental protocol of the oral administration of CLE for the cedar pollinosis model mouse. It is a graph which showed the number of times of sneezing (A) and the nasal scratching behavior (B) in the cedar pollinosis model mouse to which CLE was orally administered. It is a graph which showed the total IgE and IgG concentration in the cedar pollinosis model mouse to which CLE was orally administered. It is a graph which showed the cedar pollen antigen-specific IgE and IgG concentration in the cedar pollinosis model mouse to which CLE was orally administered.

[1] Anti-allergic agent The anti-allergic agent of the present invention contains a pulverized product or an extract of coriander. The site of use of coriander is not particularly limited, and examples thereof include whole plants, roots, stems, leaves, flowers, fruits, or seeds, or a mixture of at least two or more thereof, but leaves are preferable. Or it is a stem. Coriander may be used raw or dried (for example, freeze-dried) when performing a crushing operation or an extraction operation. In the case of extraction, it is preferable to process it into a crushed product or powder so as to improve the extraction efficiency.
Coriander is also called pakuchi in Thai, and especially the leaves that eat raw are sometimes called pakuchi.

《Crushed material》
The crushed product of coriander in the present invention is not particularly limited as long as the crushed coriander is in a crushed state, and examples thereof include powdered, granular, or paste-like crushed products, preferably powder. Is. Further, for example, a powder formed or granulated into cubes, blocks, or granules can also be preferably used. The treatment for processing into a crushed product is not particularly limited, but the plant is used by any method usually used by those skilled in the art using a crushing device or instrument such as a crusher, a mill, a blender, a mixer, and a stone mill. Examples include the process of crushing the body. The plant may be dried prior to grinding. Examples of the drying treatment method include freeze-drying, vacuum drying, blast drying, and heat-drying.

When the coriander contained in the antiallergic agent of the present invention is a pulverized product, for example, the average longest diameter of the pulverized coriander product is 0.01 to 2 mm, preferably 0.01 to 1. 5.5 mm, more preferably 0.01 to 1 mm, still more preferably 0.01 to 0.75 mm, and most preferably 0.01 to 0.5 mm can be used. Further, 90% by weight or more of the coriander pulverized product is 0.01 to 2 mm, preferably 0.01 to 1.5 mm, more preferably 0.01 to 1 mm, still more preferably 0.01 to 0.75 mm, most preferably. Preferably, one having a maximum diameter of 0.01 to 0.5 mm can be used. In addition, 90% by weight or more of the coriander crushed product is 8.6 mesh (2 mm), 10 mesh (1.7 mm), 16 mesh (1 mm), or 30 mesh (0.5 mm) in the JIS test sieve mesh conversion table. You can use the one that passes through. When the longest diameter of the coriander crushed product is 2 mm or less, the antiallergic effect of the present invention is improved, so that it is preferable to use the one having the longest diameter of 2 mm or less. The average maximum diameter of coriander can be measured using a known device for measuring the particle size. It is also possible to select arbitrarily 100 pieces from the crushed coriander, measure their longest diameters using a stereomicroscope, and calculate the average of them.

《Extract》
The extract containing the active ingredient can be extracted by a conventional extraction method used for extracting a component derived from a plant. The extraction method includes, but is not limited to, a solvent extraction method, a steam distillation method, a squeezing method (direct, high temperature or low temperature), or a supercritical extraction method. Further, a combination of these extraction methods, for example, a method of extracting with a solvent after squeezing may be used, but solvent extraction is preferable. The coriander used for extraction may be used raw or dried. Further, in order to improve the extraction efficiency, it may be processed into a crushed product or a powder before extraction.

(Solvent extraction method)
The extraction solvent used in the solvent extraction method is not limited as long as the active ingredient can be sufficiently extracted in the extract. For example, an organic solvent, an aqueous solvent, or a mixture of an organic solvent and an aqueous solvent can be used, but an aqueous solvent, or a mixture of an organic solvent and an aqueous solvent is preferable, and an aqueous solvent is more preferable.

The aqueous solvent is not limited as long as it contains water, and for example, water, physiological saline, a buffer solution, or the like can be used. Examples of the buffer solution include phosphate buffer solution, sodium phosphate buffer solution, sodium carbonate buffer solution, citrate buffer solution, citrate phosphate buffer solution, acetate buffer solution, Tris buffer solution and the like. A preferred aqueous solvent is sodium phosphate buffer. The pH of the aqueous solvent is not particularly limited.

Examples of the organic solvent include alcohol, acetone, benzene, ester, ethyl acetate, hexane, chloroform, or diethyl ether. The alcohol is not limited as long as the active ingredient can be sufficiently extracted in the extract, but for example, one of 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, isopropyl alcohol, and butyl alcohol. Examples thereof include polyhydric alcohols, 1,3-butylene glycol, propylene glycol, dipropylene glycol, and polyhydric alcohols having 2 to 5 carbon atoms such as glycerin.

The extract used for the antiallergic agent of the present invention can be extracted with a mixture of an organic solvent and an aqueous solvent. The amount of the aqueous solvent contained in the extraction solvent is not limited as long as the active ingredient can be sufficiently extracted in the extract, but the content of the aqueous solvent with respect to the total amount of the extraction solvent is, for example, , 50% by weight or more, 70% by weight or more, or 90% by weight or more.

When the extract is extracted by a solvent extraction method, the extraction temperature is not particularly limited as long as the active ingredient can be sufficiently extracted into the extract, but is -50 ° C to 100. The temperature is preferably -25 ° C to 50 ° C, more preferably -25 ° C to 25 ° C, further preferably -10 ° C to 10 ° C, and 0 ° C to 10 ° C. Most preferably, it is ° C.

Further, it is preferable to carry out the extraction while stirring or shaking so as to improve the extraction efficiency. The extraction time can be appropriately determined depending on the portion used, for example, roots, stems, leaves, flowers, fruits, or seeds. Further, the extraction time can be appropriately determined according to the coriander state, that is, the raw or dried product, or the crushed product or the powdered product, if processed. Further, the extraction time can be appropriately determined depending on the temperature of the extract or the extraction conditions such as the presence or absence of stirring or shaking. The extraction time is usually 1 minute to 72 hours, preferably 1 hour to 48 hours, and most preferably 12 hours to 36 hours.

(Steam distillation method)
The coriander extract contained in the antiallergic agent of the present invention can be extracted by a steam distillation method. The steam distillation method is a method in which steam is aerated through the raw material filled in the column, and the aroma component distilled out by the steam is condensed together with the steam. As the distillation means, any one of pressurized steam distillation, atmospheric pressure steam distillation, and reduced pressure steam distillation can be adopted.

(Squeezing method)
The squeezing method is a method of extracting an extract by physically applying pressure to coriander. There are a direct pressing method performed at room temperature, a high temperature pressing method performed at high temperature, and a low temperature pressing method performed at low temperature. The extract contained in the antiallergic agent of the present invention can be extracted by any pressing method.

(Supercritical extraction method)
The extract contained in the antiallergic agent of the present invention can be extracted by using a supercritical extraction method. The supercritical extraction method is a method of extracting an extract from a specific plant using a substance in a supercritical state. As the substance in the supercritical state, for example, carbon dioxide can be used. Since carbon dioxide in a supercritical state has a strong dissolving power, it is also generally used for decaffeination of coffee or extraction of flavors and pharmaceutical components from natural raw materials such as plants.

《Active ingredient》
The active ingredient contained in the antiallergic agent of the present invention is contained in the extract extracted from coriander. Therefore, coriander contains an ingredient having an anti-allergic effect, and the ground coriander also contains the active ingredient contained in the extract. Therefore, crushed coriander can also be used as an antiallergic agent.
The active ingredient contained in the coriander extract may be a fraction containing an active ingredient fractionated from the coriander extract or a purified active ingredient.

"allergy"
The allergy targeted by the antiallergic agent of the present invention is not particularly limited, and examples thereof include type I allergy. Type I allergies include, for example, atopic dermatitis, bronchial asthma, allergic rhinitis, pollinosis, urticaria, food allergies, animal allergies, allergic conjunctivitis, anaphylactic shock, or allergic gastroenteritis. The antiallergic agent of the present invention of the present invention has an effect of preventing the onset of allergic disease, or alleviating or treating the onset allergic disease in a subject who has developed or may develop allergic disease. ..

The type I allergic reaction involves mast cells and basophils with antigen-specific IgE antibodies and IgE-specific high-affinity IgE receptor FcεRI. When IgE bound to the surface of mast cells or basophils via FcεRI is cross-linked by an antigen, FcεRI is activated to transmit a signal downstream thereof, and histamine, leukotriene C4, PAF, Alternatively, intracellular granule contents such as eosinophil motility factor are released. This phenomenon is called degranulation. Smooth muscle contraction, increased vascular permeability, or increased glandular secretion occur in each tissue in which degranulation has occurred, and allergic symptoms appear. Elevated intracellular calcium ion concentration is also involved in degranulation. The anti-allergic agent of the present invention can inhibit the antigen-antibody reaction between the IgE antibody and the antigen. The antiallergic agent of the present invention can suppress an increase in intracellular calcium ion concentration. In addition, the antiallergic agent of the present invention can inhibit degranulation signal transduction. The antiallergic agent of the present invention may have one of these effects, or may have two or more of them.

The dosage form of the antiallergic agent of the present invention is not particularly limited, and examples thereof include oral preparations and parenteral preparations, but oral preparations are preferable. The oral preparation includes, for example, solid or powdery preparations such as fine granules, granules, tablets, capsules, and pills, and liquid preparations such as suspensions, emulsions, syrups, and extracts. Can be mentioned. Examples of parenteral preparations include injections.

The antiallergic agent of the present invention may consist of coriander pulverized product or coriander extract, or may contain coriander pulverized product or coriander extract. When the antiallergic agent of the present invention contains coriander pulverized product or coriander extract, other additives can be contained.

When the antiallergic agent of the present invention is an oral preparation, other additives include excipients, binders, disintegrants, emulsifiers, talc agents, fluidity promoters, diluents, preservatives, colorants, etc. Examples thereof include fragrances, flavoring agents, stabilizers, moisturizing agents, preservatives, antioxidants, or suspending agents, and specific examples thereof include gelatin, sodium alginate, starch, corn starch, sucrose, and lactose. Glucose, mannit, carboxymethyl cellulose, dextrin, polyvinylpyrrolidone, crystalline cellulose, soybean lecithin, sucrose, fatty acid ester, talc, magnesium stearate, polyethylene glycol, magnesium silicate, anhydrous silicic acid, synthetic aluminum silicate, etc. be able to.

When the antiallergic agent of the present invention is a parenteral agent, other additives include a water-soluble solvent such as physiological saline or Ringer's solution, a water-insoluble solvent such as vegetable oil or fatty acid ester, glucose, sodium chloride and the like. Examples thereof include tensioning agents, solubilizing agents, stabilizers, preservatives, suspending agents, emulsifiers and the like.

The antiallergic agent of the present invention can contain 90% by weight or more, 50% by weight or more, 10% by weight or more, or 1% by weight or more of coriander pulverized product or extract.

The dose or intake of the antiallergic agent of the present invention can be appropriately adjusted according to the form of the preparation and the age, sex, body weight, degree of allergic symptoms, etc. of the subject to be used, and the antiallergic agent is administered. Alternatively, the amount is preferably such that the onset of allergic disease can be prevented, or the allergic disease that has developed can be alleviated or treated by ingestion. Specifically, in terms of the amount of the coriander aqueous solvent extract added, 0.01 to 1000 mg / kg body weight / day, preferably 0.1 to 750 mg / kg body weight / day, more preferably 1 to 500 mg / day. kg body weight / day, more preferably 5 to 400 mg / kg body weight / day, still more preferably 10 to 300 mg / kg body weight / day, still more preferably 15 to 200 mg / kg body weight / day, or most preferably 20 to 150 mg / kg. Can be weight / day. Of course, the above administration method is an example, and other administration methods may be used. It is desirable that the method, dose, administration period, administration interval, etc. of the antiallergic agent to humans be determined by a controlled clinical trial.

The antiallergic agent of the present invention can be administered to humans, but the administration target may be animals other than humans, and pets such as dogs, cats, rabbits, hamsters, guinea pigs, and squirrels; cows and animals. Livestock such as pigs; experimental animals such as mice and rats; and animals bred in zoos and the like can be mentioned.

The antiallergic agent of the present invention can be a pharmaceutical composition for preventing or treating allergies. The pharmaceutical composition includes pharmaceuticals and quasi-drugs. Examples of the pharmaceutical product include crude drug preparations and Chinese herbal preparations. Examples of quasi-drugs include energy drinks and herbal medicine-containing health medicines.

[2] Anti-allergic food composition In the present specification, the food composition means a food or a beverage containing the anti-allergic agent of the present invention. The food composition of the present invention contains the antiallergic agent of the present invention and can therefore be used as a food composition for preventing or treating allergies.

Specific foods include fresh cooked foods such as salads; cooked foods such as steaks, pizzas and hamburgers; fried cooked foods such as fried vegetables; tomatoes, peppers, celery, niggauri, carrots, potatoes and asparagus. Vegetables such as and cooked products processed from these vegetables; cookies, bread, biscuits, dried bread, cakes, rice cakes, sheep rice, pudding, jelly, ice creams, chewing gum, crackers, chips, chocolate and sweets such as candy; udon, Noodles such as pasta and buckwheat; Fish paste products such as kamaboko, ham, and fish sausage; Dairy products such as cheese, cream, and butter; Miso, soy sauce, dressing, ketchup, mayonnaise, soup base, noodle soup, curry powder , Seasonings such as mirin and ru; soy foods such as tofu; processed agricultural and marine products such as sprinkles, boiled vegetables and cereals; and konjac.

Beverages include, for example, coffee beverages; cocoa beverages; vegetable juices obtained from the above vegetables; fruit juice beverages such as grapefruit juice, orange juice, grape juice, and lemon juice; tea beverages such as green tea, tea, roasted tea, and oolong tea. Alcoholic beverages such as beer, wine (red wine, white wine, or sparkling wine), sake, plum wine, sparkling liquor, whiskey, brandy, shochu, lamb, gin, and liqueurs; milk beverage; soy milk beverage; liquid food; In addition, sports drinks and the like can be mentioned.

Foods or beverages include feeds and beverages for animals. Examples of the target animal include primates such as humans, cows, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, mice and the like.

These foods or beverages may optionally include antioxidants, fragrances, acidulants, colorants, emulsifiers, preservatives, seasonings, sweeteners, spices, pH regulators, stabilizers, vegetable oils, animal oils, sugars and sugars. Food additives and food materials such as alcohols, vitamins, organic acids, fruit juice extracts, vegetable extracts, grains, beans, vegetables, meats, seafood, etc. can be blended alone or in combination of two or more. The blending amount of these food materials and food additives can be appropriately determined within a range that does not impair the object of the present invention.

These foods or beverages are commonly used, for example, in heat and pressure sterilization such as retort and autoclave, batch sterilization, plate sterilization, energization heat sterilization, microwave heat sterilization, and steam sterilization such as injection and infusion. Can be sterilized.

Foods and beverages include functional foods (beverages) and health foods (beverages). In the present specification, the "health food (beverage)" means a food or beverage that has or can be expected to have some effect on health, and the "functional food (beverage)" is described above. Among "health foods (beverages)", foods designed and processed so as to sufficiently exert a bioregulatory function (that is, a function of preventing the onset of allergic symptoms or a function of alleviating or treating allergic symptoms). Means beverage. Functional foods and health foods can be in the form of granules, solids, liquids, capsules, gels or tablets.

Hereinafter, the present invention will be specifically described with reference to Examples, but these do not limit the scope of the present invention.

<< Preparation Example 1 >>
In this preparation example, an extract was prepared from fresh coriander (pakuchi) leaves using sodium phosphate buffer. The adjustment method is summarized in FIG.
Fresh leaves of coriander were chopped and freeze-dried to make a powder. Pakuchi leaf lyophilized powder was suspended in 10 mM sodium phosphate buffer (NaPB; pH 7.4) at 0.05 g / mL and extracted at 4 ° C. for 24 hours. Then, the insoluble material was removed by centrifugation, and the mixture was sterilized by filtration using a 0.45 μm filter.
In order to prevent the endogenous enzyme in the sample from affecting the test for antiallergic effect, the endogenous enzyme was inactivated by heat treatment at 100 ° C. for 5 minutes. The obtained coriander leaf water-soluble extract may be referred to as CLE.

<< Example 1: Degranulation test >>
In this example, the obtained CLE was subjected to a degranulation test.
Rat basal cell line RBL-2H3 cells were precultured in ⁵ % FBS-DMEM medium in 96-well culture plates at a cell concentration of 2 × 105 cells / mL for 18 hours. The cells were washed once with phosphate buffered saline (PBS). It was then sensitized with dinitrophenyl (DNP) specific IgE for 2 hours. The cells were washed twice with Tyrode buffer. A sample of cumin water-soluble extract diluted with Tyrode buffer was added at 200 μL / well and cultured for 10 minutes. After 10 minutes, the cilantro water-soluble extract sample solution diluted with Tyrode buffer was aspirated and discarded, and Tyrode buffer was added at 200 μL / well. Subsequently, the antigen DNP was added and cultured for 30 minutes to induce degranulation, and then the supernatant was collected.

The degranulation inhibitory effect of CLE was evaluated using the amount of β-hexosaminidase released in the granules released by antigen stimulation as an index. Specifically, after collecting the supernatant, the cells were sonicated on ice for 5 seconds in 130 μL of modified Tyrode buffer containing 0.1% Triton X-100 to lyse the cells. Both the supernatant recovered above and the cytolysate obtained above were transferred to each well of a new 96-well (50 μL / well) microplate and incubated at 37 ° C. for 5 minutes. Then, 100 μL of 3.3 mM 4-nitrophenyl2-acetamide-2-deoxy-β-D-glucopyranoside (substrate solution) dissolved in 0.1 M citrate buffer (pH 4.5) was added to each well. , Further incubated at 37 ° C. for 25 minutes. The enzymatic reaction was stopped by adding 100 μL of 2M glycine buffer (pH 10.4). The absorbance of the reaction solution was measured at a wavelength of 405 nm using a microplate reader (SH-8000Lab, Corona Electric). The β-hexosaminidase release rate (%) was calculated according to the following formula as an index of degranulation.

β-hexosaminidase release rate (%) = [(A _supernatant -A _{supernatant blank} ) / {(A _supernatant -A _{supernatant blank} ) + (A _{cell lysate-} A cell _lysate blank) )}] × 100

A: Absorbance of each well at a wavelength of 405 nm

The protein concentration was measured according to the Lowry method. Specifically, it was quantified using a DC protein assay kit manufactured by Bio-Rad.

As shown in FIG. 2 (A), the coriander leaf water-soluble extract suppressed degranulation of RBL-2H3 cells in a concentration-dependent manner.
Moreover, when the cytotoxicity of the Pakuchi leaf water-soluble extract was measured by the WST-8 method, as shown in FIG. 2 (B), the Pakuchi leaf water-soluble extract showed cytotoxicity at a concentration that suppresses degranulation. There wasn't.
In addition, the coriander leaf water-soluble extract was dialyzed against dialysis membranes having a molecular weight cut-off of 14,000 and 500, and the effect of suppressing degranulation was measured. As a result, treatment with 14,000 and 500 dialysis membranes reduced activity. Therefore, it was suggested that the small molecule fraction with a molecular weight of 500 or less may contain at least a part of the component that suppresses degranulation.

<< Example 2 >>
In this example, changes in intracellular calcium ion concentration due to coriander leaf water-soluble extract (CLE) were measured. CLE with a protein concentration of 4000 μg / mL was added to RBL-2H3 cells, and the intracellular calcium ion concentration before and after stimulation with dinitrophenyl (DNP) -specific IgE was measured over time. As a result, as shown in FIG. 3, the increase in intracellular calcium ion concentration was suppressed by the addition of CLE.

<< Example 3 >>
In this example, the inhibitory effect of CLE on degranulation by A23187 stimulation was examined.
The operation of Example 1 was repeated except that stimulation with A23187 (1 μM) was performed instead of stimulation with dinitrophenyl (DNP) -specific IgE. As shown in FIG. 4 (A), CLE suppressed degranulation caused by A23187 stimulation in a concentration-dependent manner.
In addition, except that the stimulation with A23187 was used instead of the stimulation with dinitrophenyl (DNP) -specific IgE, and the protein concentration of CLE was 1000 μg / mL, 2000 μg / mL, and 4000 μg / mL, the examples were used. The operation of 2 was repeated, and the intracellular calcium concentration was measured. As shown in FIG. 4 (B), CLE suppressed the increase in intracellular calcium concentration caused by A23187 stimulation.

<< Example 4 >>
In this example, the inhibitory effect of CLE on degranulation by Thapsigargin stimulation was investigated.
The operation of Example 3 was repeated except that the stimulation with Thapsigargin (1 μM) was performed instead of the stimulation with A23187. As shown in FIGS. 5A and 5B, CLE suppressed degranulation caused by Thapsigargin stimulation in a concentration-dependent manner, and suppressed an increase in intracellular calcium concentration.

<< Example 5 >>
In this example, the effect of coriander leaf water-soluble extract on degranulation signaling was investigated.
After adding CLE (4000 μg / mL) and culturing RBL-2H3 cells, a cell disruption solution was prepared and subjected to SDS-PAGE gel electrophoresis. After electrophoresis, the electrophoresed protein was transferred to the PVDF membrane by Western blotting. The transcribed PVDF membrane was blocked with skim milk and then reacted with each signal factor-specific antibody (manufactured by Cell Signaling Technology). After reacting with a peroxidase-labeled anti-IgG antibody, the substrate solution was reacted and a band was detected.
As a result, as shown in FIG. 6, it was found that CLE suppresses the activation of PI3K protein by phosphorylation. Further, as shown in FIG. 7, PLCγ and Akt are downstream factors of PI3K, but it is considered that activation of PLCγ and Akt is also suppressed by suppressing phosphorylation of PI3K (FIG. 6). ).
From these results, it was speculated that CLE may inhibit both Ca ²⁺ concentration-dependent pathway and non-Ca 2+ concentration-dependent pathway by suppressing the activation level of PK3I.

<< Example 7 >>
In this example, the effect of CLE on Japanese cedar pollinosis model mice was examined.
The effect of oral administration of CLE on mice that developed cedar pollinosis after antigen sensitization with cedar pollinosis antigens (Cryj1 and Cryj2) was investigated. The experimental protocol is shown in FIG. Stimulation was performed 7 times in a row on a 2.0 μg cedar pollen plateau for 7 consecutive days. During 7 days of antigen stimulation, CLE was orally administered at two concentrations of 50 mg / kg body weight / day (low dose) and 250 mg / kg body weight / day (high dose), and the effect on the symptoms was examined. Blood antibodies were also measured.

As a result of measuring the number of times of sneezing for 15 minutes after the last antigen stimulation, a remarkable symptom-improving effect was observed in the high-dose administration group (Fig. 9A). On the other hand, as a result of similarly measuring the scratching behavior of the nose, it was clarified that the symptoms were alleviated in the high-dose group to the same extent as in the non-sensitized group, although there was no significant difference (Fig. 9B).

Moreover, as a result of measuring the antigen-nonspecific IgE and IgG1 concentrations in blood, it was clarified that both IgE and IgG were remarkably suppressed (FIG. 10). In addition, as a result of measuring blood IgE and IgG1 specific to cedar pollen antigen, although it was not significant, the IgE concentration tended to be suppressed, and the IgG concentration was significantly suppressed in the low-dose administration group (FIG. 11). ).

Claims (2)

An antiallergic agent for type I allergies (excluding food allergies), which comprises a crushed coriander leaf or an extract of an aqueous solvent or a mixture of an aqueous solvent and an organic solvent . A food composition for preventing or treating an allergic disease, which comprises the antiallergic agent according to claim 1 and a food or drink.

Images